PCR detection,quantitative real-time PCR(q-RTPCR),outdoor insect resistance,and disease resistance identification were carried out for the detection of genetic stability and disease resistance through generations(T2,T...PCR detection,quantitative real-time PCR(q-RTPCR),outdoor insect resistance,and disease resistance identification were carried out for the detection of genetic stability and disease resistance through generations(T2,T3,and T4)in transgenic maize germplasms(S3002 and 349)containing the bivalent genes(insect resistance gene Cry1Ab13-1 and disease resistance gene NPR1)and their corresponding wild type.Results indicated that the target genes Cry1Ab13-1 and NPR1 were successfully transferred into both germplasms through tested generations;q-PCR confirmed the expression of Cry1Ab13-1 and NPR1 genes in roots,stems,and leaves of tested maize plants.In addition,S3002 and 349 bivalent gene-transformed lines exhibited resistance to large leaf spots and corn borer in the field evaluation compared to the wild type.Our study confirmed that Cry1Ab13-1 and NPR1 bivalent genes enhanced the resistance against maize borer and large leaf spot disease and can stably inherit.These findings could be exploited for improving other cultivated maize varieties.展开更多
背景与目的热休克蛋白90AB1(heat shock protein 90 k Da alpha,class B member 1,Hsp90AB1)是ATP依赖的高度保守的分子伴侣,在多种肿瘤细胞中过表达。在肿瘤发生发展的信号传导通路中起着重要作用的一些分子,如表皮生长因子受体(epider...背景与目的热休克蛋白90AB1(heat shock protein 90 k Da alpha,class B member 1,Hsp90AB1)是ATP依赖的高度保守的分子伴侣,在多种肿瘤细胞中过表达。在肿瘤发生发展的信号传导通路中起着重要作用的一些分子,如表皮生长因子受体(epidermal growth factor receptor,EGFR)、人类表皮生长因子受体-2(human epidermal growth factor receptor-2,HER2)等,均是Hsp90AB1的底物蛋白。Hsp90AB1与这些底物蛋白相互作用并参与细胞的多种病理生理过程。本研究通过检测Hsp90AB1在非小细胞肺癌(non-small cell lung cancer,NSCLC)组织中的蛋白表达情况以初步探讨其临床意义。方法采用组织微阵列和免疫组织化学染色的方法检测Hsp90AB1在213例NSCLC及相应癌旁正常肺组织中的蛋白表达,并分析Hsp90AB1的表达与NSCLC临床病理参数及患者预后的关系。结果 Hsp90AB1在肺癌组织中的表达水平(阳性率54.0%)高于在正常肺组织中的表达水平(阳性率0.0%,P<0.001)。Hsp90AB1在肺腺癌中的表达阳性率为61.2%,高于肺鳞癌组织37.9%(P=0.002),并且其高表达与肺腺癌患者的不良预后相关(P=0.032)。Hsp90AB1蛋白表达水平与临床分期、淋巴结转移、病理分级等因素无关(P>0.05)。结论 Hsp90AB1在NSCLC组织中高表达,并且其表达水平与肺癌的病理类型及肺腺癌患者总生存期相关。展开更多
【目的】构建包含抗鳞翅目害虫基因cry1Ab13的重组植物表达载体,并利用其创制对玉米螟Ostrinia furnacalis具有优良抗性的转基因玉米Zea may L.新种质。【方法】利用同源重组法将cry1Ab13基因连接到表达载体pCAMBIA3300-Bar上,获得以抗...【目的】构建包含抗鳞翅目害虫基因cry1Ab13的重组植物表达载体,并利用其创制对玉米螟Ostrinia furnacalis具有优良抗性的转基因玉米Zea may L.新种质。【方法】利用同源重组法将cry1Ab13基因连接到表达载体pCAMBIA3300-Bar上,获得以抗除草剂Bar基因为筛选标记的植物表达载体pCAMBIA3300-cry1Ab13-Bar。通过农杆菌介导法转化玉米自交系H99幼胚,对再生植株进行逐代除草剂筛选、PCR检测及T2代植株的Southern blotting检测、实时荧光定量PCR检测,并对转基因植株进行田间及室内抗虫性鉴定。【结果】构建了cry1Ab13基因的植物表达载体,转化玉米获得3株高抗玉米螟和1株抗玉米螟的T2代转基因植株。Southern blotting证明cry1Ab13基因已经整合到玉米基因组中,实时荧光定量PCR结果表明cry1Ab13基因已经在玉米植株内表达。抗虫性鉴定结果表明,与对照相比转基因植株对玉米螟的抗性显著提高。【结论】将cry1Ab13基因导入玉米并成功表达,显著提高了转基因玉米对玉米螟的抗性,为抗虫玉米新种质的创制奠定了基础。展开更多
As a highly important fish virus,nervous necrosis virus(NNV)has caused severe economic losses to the aquaculture industry worldwide.Autophagy,an evolutionarily conserved intracellular degradation process,is involved i...As a highly important fish virus,nervous necrosis virus(NNV)has caused severe economic losses to the aquaculture industry worldwide.Autophagy,an evolutionarily conserved intracellular degradation process,is involved in the pathogenesis of several viruses.Although NNV can induce autophagy to facilitate infection in grouper fish spleen cells,how it initiates and mediates autophagy pathways during the initial stage of infection is still unclear.Here,we found that red-spotted grouper NNV(RGNNV)induced autophagosome formation in two fish cell lines at 1.5 and 3 h post infection,indicating that autophagy is activated upon entry of RGNNV.Moreover,autophagic detection showed that RGNNV entry induced incomplete autophagy by impairing the fusion of autophagosomes with lysosomes.Further investigation revealed that binding of the RGNNV capsid protein(CP)to the Lateolabrax japonicus heat shock protein HSP90ab1(LjHSP90ab1),a cell surface receptor of RGNNV,contributed to RGNNV invasion-induced autophagy.Finally,we found that CP blocked the interaction of L.japonicus protein kinase B(AKT)with LjHSP90ab1 by competitively binding the NM domain of LjHSP90ab1 to inhibit the AKT-mechanistic target of the rapamycin(MTOR)pathway.This study provides novel insight into the relationship between NNV receptors and autophagy,which may help clarify the pathogenesis of NNV.展开更多
基金supported by the National Key Research and Development Program of China(2019YFD1002603-1)。
文摘PCR detection,quantitative real-time PCR(q-RTPCR),outdoor insect resistance,and disease resistance identification were carried out for the detection of genetic stability and disease resistance through generations(T2,T3,and T4)in transgenic maize germplasms(S3002 and 349)containing the bivalent genes(insect resistance gene Cry1Ab13-1 and disease resistance gene NPR1)and their corresponding wild type.Results indicated that the target genes Cry1Ab13-1 and NPR1 were successfully transferred into both germplasms through tested generations;q-PCR confirmed the expression of Cry1Ab13-1 and NPR1 genes in roots,stems,and leaves of tested maize plants.In addition,S3002 and 349 bivalent gene-transformed lines exhibited resistance to large leaf spots and corn borer in the field evaluation compared to the wild type.Our study confirmed that Cry1Ab13-1 and NPR1 bivalent genes enhanced the resistance against maize borer and large leaf spot disease and can stably inherit.These findings could be exploited for improving other cultivated maize varieties.
文摘背景与目的热休克蛋白90AB1(heat shock protein 90 k Da alpha,class B member 1,Hsp90AB1)是ATP依赖的高度保守的分子伴侣,在多种肿瘤细胞中过表达。在肿瘤发生发展的信号传导通路中起着重要作用的一些分子,如表皮生长因子受体(epidermal growth factor receptor,EGFR)、人类表皮生长因子受体-2(human epidermal growth factor receptor-2,HER2)等,均是Hsp90AB1的底物蛋白。Hsp90AB1与这些底物蛋白相互作用并参与细胞的多种病理生理过程。本研究通过检测Hsp90AB1在非小细胞肺癌(non-small cell lung cancer,NSCLC)组织中的蛋白表达情况以初步探讨其临床意义。方法采用组织微阵列和免疫组织化学染色的方法检测Hsp90AB1在213例NSCLC及相应癌旁正常肺组织中的蛋白表达,并分析Hsp90AB1的表达与NSCLC临床病理参数及患者预后的关系。结果 Hsp90AB1在肺癌组织中的表达水平(阳性率54.0%)高于在正常肺组织中的表达水平(阳性率0.0%,P<0.001)。Hsp90AB1在肺腺癌中的表达阳性率为61.2%,高于肺鳞癌组织37.9%(P=0.002),并且其高表达与肺腺癌患者的不良预后相关(P=0.032)。Hsp90AB1蛋白表达水平与临床分期、淋巴结转移、病理分级等因素无关(P>0.05)。结论 Hsp90AB1在NSCLC组织中高表达,并且其表达水平与肺癌的病理类型及肺腺癌患者总生存期相关。
基金supported by the Pearl River S&T Nova Program of Guangzhou(201806010047)National Natural Science Foundation of China(32173001,3210284,31771587)+1 种基金China Postdoctoral Science Foundation Funded Project(2021M693678)Natural Science Foundation of Guangxi Province(2021GXNSFDA075015)。
文摘As a highly important fish virus,nervous necrosis virus(NNV)has caused severe economic losses to the aquaculture industry worldwide.Autophagy,an evolutionarily conserved intracellular degradation process,is involved in the pathogenesis of several viruses.Although NNV can induce autophagy to facilitate infection in grouper fish spleen cells,how it initiates and mediates autophagy pathways during the initial stage of infection is still unclear.Here,we found that red-spotted grouper NNV(RGNNV)induced autophagosome formation in two fish cell lines at 1.5 and 3 h post infection,indicating that autophagy is activated upon entry of RGNNV.Moreover,autophagic detection showed that RGNNV entry induced incomplete autophagy by impairing the fusion of autophagosomes with lysosomes.Further investigation revealed that binding of the RGNNV capsid protein(CP)to the Lateolabrax japonicus heat shock protein HSP90ab1(LjHSP90ab1),a cell surface receptor of RGNNV,contributed to RGNNV invasion-induced autophagy.Finally,we found that CP blocked the interaction of L.japonicus protein kinase B(AKT)with LjHSP90ab1 by competitively binding the NM domain of LjHSP90ab1 to inhibit the AKT-mechanistic target of the rapamycin(MTOR)pathway.This study provides novel insight into the relationship between NNV receptors and autophagy,which may help clarify the pathogenesis of NNV.
